PROJECT SUMMARY - GENE EXPRESSION AND MANIPULATION The Gene Expression and Manipulation Core is focused on providing assistance and facilitating studies relating to evaluating, quantitatively, the temporal effects of treatments, conditions or disease states on the expression levels, mRNA and protein, of genes in ocular tissues. It is also focused on determining the effects of individual gene manipulation on ocular processes and cells. This includes knockdown of specific gene expression levels in cells or tissues by RNAi silencing, increases in specific gene expression by overexpression, and modulation of activity or function of specific genes by expressing modified forms, e.g. by transfecting or infecting genetic mutant, dominant negative, constitutively active or alternatively spliced gene constructs. Manipulation methods include construct transfection and viral gene or RNAi delivery via infection with recombinant viral constructs. Expression studies using PCR Arrays and cis element promoter studies are also facilitated. We have and maintain instrumentation for these types of studies. We have experience with all of these types of methods and will facilitate and assist core users with any of these that they deem of value to their investigations. In addition to providing this service to NEI funded investigators, we actively support efforts for investigators, particularly new and young investigators, to develop pilot or preliminary data that can be leveraged into future foundation and NEI funding.